畜牧与兽医2024，Vol.56Issue(4)：66-71,6.
阪崎克罗诺杆菌双抗夹心酶联免疫吸附检测方法的建立
Establishment of a double-antibody sandwich enzyme-linked immunosorbent assay for Cronobacter sakazakii detection
摘要
Abstract
To establish a method for rapid immunological detection of Cronobacter sakazakii,a double antibody sandwich enzyme-linked immunosorbent assay was tried in this study.The representative strains of C.sakazakii were used as mixed antigens to immunize mice.Four strains of monoclonal antibodies(McAbs)against C.sakazakii were prepared by the hybridoma technique,with the titers being 1 ∶ 128 000,1 ∶ 256 000,1 ∶ 32 000 and 1 ∶ 256 000,respectively.The titer of polyclonal antibodies(PcAbs)titers was 1 ∶ 128 000.To establish the double antibody sandwich ELISA,the anti-C.sakazakii PcAbs was used as the capture antibodies and anti-C.sakazakii McAbs as detecting antibodies,respectively.The optimum dilution concentrations of PcAbs and McAbs were 1 ∶ 4 000 and 1 ∶ 1 000,respectively.The sensitivi-ty of this method was 1×106 CFU/mL,and the method had good specificity with no cross-reactivity with 6 other foodborne pathogens,and it could simultaneously detect six Cronobacter species.The results of the repetitive test showed that the intra-and inter-batch coefficients of variation of the method were less than 15%.In summary,a double-antibody sandwich ELISA method was successfully established here using anti-C.sakazakii McAbs and anti-C.sakazakii PcAbs.This method possessed good sensitivity,specificity,and reproducibility;which would be a technical support for rapid screening and analysis of C.sakazakii.关键词
阪崎克罗诺杆菌/单克隆抗体/双抗体夹心ELISAKey words
Cronobacter sakazakii/monoclonal antibody/double-antibody sandwich ELISA分类
农业科技引用本文复制引用
张欣,王福成,韩先干,祁克宗,宋祥军,董雨豪,蒋蔚..阪崎克罗诺杆菌双抗夹心酶联免疫吸附检测方法的建立[J].畜牧与兽医,2024,56(4):66-71,6.基金项目
上海市科技兴农重点攻关项目(2020-02-08-00-08-F01489) （2020-02-08-00-08-F01489）
上海市科学技术委员会科研计划项目(21N31901000) （21N31901000）
国家重点研发计划政府间国际科技创新合作重点专项(2018YFE0102200) （2018YFE0102200）
兵团财政科技计划项目(2020AB025-03) （2020AB025-03）
中央级公益性科研院所基本科研业务费专项资金项目(2020JB07) （2020JB07）
